Mechanism and Anticancer Activity of the Metabolites of an Endophytic Fungi from Eucommia ulmoides Oliv.

Backgroud: Pinoresinol (Pin) and pinoresinol monoglucoside (PMG) are plant-derived lignan molecules with multiple functions. We showed previously that an endophytic fungus from Eucommia ulmoides Oliv., Phomopsis sp. XP-8 is able to produce Pin and PMG. OBJECTIVE: This study was carried out to test the anti-tumor capability of the culture of XP-8 and identify the major effective compounds. METHOD: The fungal culture was added in the culture of HepG2 and K562 cells, and the viabilities of these cells were detected and the possible mechanism was analyzed. RESULT: The fungal culture showed significant capaiblity in decreasing the viability of tumor cells and induce apoptosis via up-regulation of the expression of apoptosis-related genes. It also significantly inhibited the adhesion and migration of HepG2 cells by blocking MMP-9 expression. Pin and PMG were isolated from the growth culture and shown to be the major effective components for inhibition. CONCLUSION: The study indicated the potential application of XP-8 in the production of anti-tumour products by the bioconversion of glucose.

HIRA is essential for the development of gibel carp.

HIRA is one of the chaperones of histone H3.3. Mutation of Hira results in embryonic lethality in mice, suggesting a critical role in embryogenesis. However, Hira-mutated Drosophila may survive to adults, indicating that it is dispensable in Drosophila development. The role of Hira in fish development is unknown. In this study we first investigated the expression of Hira during embryogenesis of gibel carp (Carassius auratus gibelio) by whole-mount in situ hybridization. We found that Hira signal appeared ubiquitously in the early embryos. After gastrulation, it appeared mainly along the anterior-posterior axis, including the tail bud. In hatching period, the signal was detected in head, heart, and the endoderm region on the back of yolk. Then by microinjection with morpholino-HIRA at the beginning of development, we observed delayed gastrulation and abnormal somitogenesis in gibel carp embryos. The HIRA morphants exhibited short trunk, limited yolk extension, and twisted tail. Most of the mutants died during embryogenesis or shortly after hatching. The rest of the HIRA morphants could survive to larvae but with severe defects in organogenesis. These data suggest that HIRA may be essential for the development of gibel carp, and this function is conserved in vertebrates.

Expression profiles of six novel C-type lectins in response to bacterial and 20E injection in the cotton bollworm (Helicoverpa armigera).

C-type lectins can act as pattern recognition receptors (PRRs) and play an important role in innate immunity. Two C-type lectins (HaCTL1 and HaCTL2) have been previously identified in the cotton bollworm (Helicoverpa armigera). Here we isolate six C-type lectins from H. armigera (HaCTL3, 4, 5, 6, 7 and 8). All six new HaCTLs encode a signal peptide (or partial signal peptide) and complete tandem carbohydrate-recognition domains (CRDs). HaCTL4, 5, 6, 7 and 8 mRNA increased in the fat body after injection with both killed Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus, whereas HaCTL3 mRNA was upregulated following E. coli injection only. Recombinant HaCTL3 exhibited agglutinating activity against both Gram-negative and Gram-positive bacteria in a calcium-dependent manner. Agglutination inhibitory analysis indicated that rHaCTL3 recognizes maltose, trehalose, peptidoglycan and lipopolysaccharides. HaCTL3 and HaCTL8 mRNA showed upregulation while HaCTL4, 5, and 6 mRNA downregulation post 20-Hydroxyecdysone (20E) injection. Our results indicate that the six novel C-type lectins of H. armigera may play important roles in defending against bacteria as PRRs and the hormone 20E can function in regulating immunity through lectins.

[Establishment and identification of highly expressing and replicating hepatitis B virus genome transgenic mouse models].

OBJECTIVE: To establish a highly expressing and replicating hepatitis B virus (HBV) genome transgenic mouse models for screening anti-HBV drugs and investigating the pathogenesis of hepatitis B. METHODS: Elongated HBV genome as the investigated gene was transducted into the pronuclei of the fertilized eggs of mice by the technique of microinjection, then the eggs were transplanted into the oviducts of the pseudopregnant mice. All the newborn mice were screened and identified by PCR and Southern blot detecting genomic DNA in tail tissue, then the positive mice were examined plasma HBsAg, HBeAg by ELISA and plasma HBV DNA by Southern blot. RESULTS: Among the 61 offsprings, 18 were positive for tail tissue HBV DNA examination, 7 of which were positive for replication and expression detection. CONCLUSION: Transgenic mice with elongated HBV genome possess high efficiency of replication and expression, which can be used for further investigation.

Nuclear transplantation of somatic cells of transgenic red carp (Cyprinus carpio haematopterus).

Nuclear transplantation was performed using kidney cells, tail fin cells and cultured tail fin cells of 18th passage of the F4 hGH gene transferred red carp (Cyprinus carpio haematopterus) as donors and the eggs of loach (Misgurnus anguillicaudatus) of Huanghe carp (Cyprinus carpio haematopterus) as recipients. Using loach unfertilized eggs as recipients 0.33% of reconstructed embryos developed in to neural plate from kidney cells, and 0.1% of reconstructed embryos developed in to muscular reaction stage from tail fin cells of F4hGH gene transferred red carp. A tatal of 0.4% reconstructed embryos from cultured tail fin cells of 18th passage and unfertilized eggs of Huanghe carp developed in to myotome stage. All the nuclear transfer embryos were positive for hGH gene by PCR analysis. The results suggest that exogenous hGH gene is stable in the F4hGH gene transferred red carp, hGH gene exists in all the cells of the F4 transgenic red carp. Somatic cell nuclear transfer is an effective pathway for producing homogenous transgenic fish, but with low efficiency. The reasons of low efficiency of somatic nuclear transplantation may be incompatibility of nuclear and ooplasm, incongruity of cell cycle, aberrance of chromosomes, etc.